scholarly journals A novel R2R3-MYB from grape hyacinth, MaMybA, which is different from MaAN2, confers intense and magenta anthocyanin pigmentation in tobacco

2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Kaili Chen ◽  
Lingjuan Du ◽  
Hongli Liu ◽  
Yali Liu
Keyword(s):  
Anthocyanin Biosynthesis ◽  
Bimolecular Fluorescence Complementation ◽  
Luciferase Assay ◽  
Myb Transcription Factor ◽  
Bhlh Protein ◽  
Anthocyanin Pigmentation ◽  
High Concentration ◽  
Shared Identity ◽  
R2r3 Myb ◽  
And Control

Abstract Background The primary pigments in flowers are anthocyanins, the biosynthesis of which is mainly regulated by R2R3-MYBs. Muscari armeniacum is an ornamental garden plant with deep cobalt blue flowers containing delphinidin-based anthocyanins. An anthocyanin-related R2R3-MYB MaAN2 has previously been identified in M. armeniacum flowers; here, we also characterized a novel R2R3-MYB MaMybA, to determine its function and highlight similarities and differences between MaMybA and MaAN2. Results In this study, a novel anthocyanin-related R2R3-MYB gene was isolated from M. armeniacum flowers and functionally identified. A sequence alignment showed that MaMybA contained motifs typically conserved with MaAN2 and its orthologs. However, the shared identity of the entire amino acid sequence between MaMybA and MaAN2 was 43.5%. Phylogenetic analysis showed that they were both clustered into the AN2 subgroup of the R2R3-MYB family, but not in the same branch. We also identified a IIIf bHLH protein, MabHLH1, in M. armeniacum flowers. A bimolecular fluorescence complementation assay showed that MabHLH1 interacted with MaMybA or MaAN2 in vivo; a dual luciferase assay indicated that MaMybA alone or in interaction with MabHLH1 could regulate the expression of MaDFR and AtDFR, but MaAN2 required MabHLH1 to do so. When overexpressing MaMybA in Nicotiana tabacum ‘NC89’, the leaves, petals, anthers, and calyx of transgenic tobacco showed intense and magenta anthocyanin pigments, whereas those of OE-MaAN2 plants had lighter pigmentation. However, the ovary wall and seed skin of OE-MaMybA tobacco were barely pigmented, while those of OE-MaAN2 tobacco were reddish-purple. Moreover, overexpressing MaMybA in tobacco obviously improved anthocyanin pigmentation, compared to the OE-MaAN2 and control plants, by largely upregulating anthocyanin biosynthetic and endogenous bHLH genes. Notably, the increased transcription of NtF3′5′H in OE-MaMybA tobacco might lead to additional accumulation of delphinidin 3-rutinoside, which was barely detected in OE-MaAN2 and control plants. We concluded that the high concentration of anthocyanin and the newly produced Dp3R caused the darker color of OE-MaMybA compared to OE-MaAN2 tobacco. Conclusion The newly identified R2R3-MYB transcription factor MaMybA functions in anthocyanin biosynthesis, but has some differences from MaAN2; MaMybA could also be useful in modifying flower color in ornamental plants.

scholarly journals A MYB transcription factor, PlMYB308, plays an essential role in flower senescence of herbaceous peony

2021 ◽  
Author(s):  
Xiaotong Ji ◽  
Zhuangzhuang Xu ◽  
Meiling Wang ◽  
Xuyang Zhong ◽  
Lingling Zeng ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Anthocyanin Biosynthesis ◽  
Bimolecular Fluorescence Complementation ◽  
Luciferase Assay ◽  
Flower Senescence ◽  
Myb Transcription Factor ◽  
Vase Life ◽  
Herbaceous Peony ◽  
Expression Levels ◽  
Variable Expression

AbstractHerbaceous peony is an important cut-flower plant cultivated across the world, but its short vase life substantially restricts the economic value of this crop. It is well established that endogenous hormones regulate the senescing process, but the molecular mechanism of them in flower senescence is still unclear. Here, we isolated a MYB transcription factor gene PlMYB308 from herbaceous peony flowers. Transcript abundance of PlMYB308 was strongly up-regulated in senescing petals. Silencing of PlMYB308 resulted in delayed peony flower senescence, and dramatically increased gibberellin (GA) but reduced ethylene and abscisic acid (ABA) levels in petals. Ectopic overexpression of PlMYB308 in tobacco accelerated flower senescence, and reduced GA but increased ethylene and ABA accumulation. Correspondingly, biosynthetic genes of ethylene, ABA, and GA showed variable expression levels in petals after silencing or overexpression of PlMYB308. A dual-luciferase assay showed that PlMYB308 specifically bound to the PlACO1 promoter. High expression levels of PlMYB308 were accompanied by low petal anthocyanin accumulation in senescing petals. A further bimolecular fluorescence complementation assay revealed an interaction between PlMYB308 and PlbHLH33, which was supposed to inhibit the anthocyanin biosynthesis. Taken together, our results suggest that the PlMYB308-PlACO1 and PlMYB308-PlbHLH33 regulatory checkpoints perhaps positively and negatively operate the production of ethylene and anthocyanin, respectively, and thus contribute to the senescence with impaired pigmentation in herbaceous peony flowers.

scholarly journals The MIR-Domain of PbbHLH2 Is Involved in Regulation of the Anthocyanin Biosynthetic Pathway in ”Red Zaosu” (PyrusBretschneideri Rehd.) Pear Fruit

2021 ◽  
Vol 22 (6) ◽  
pp. 3026
Author(s):  
Xieyu Li ◽  
Fangxin Xiang ◽  
Wei Han ◽  
Bingqing Qie ◽  
Rui Zhai ◽  
...  
Keyword(s):  
Biosynthetic Pathway ◽  
Anthocyanin Biosynthesis ◽  
Bimolecular Fluorescence Complementation ◽  
Anthocyanin Content ◽  
Pear Fruit ◽  
Fruit Peel ◽  
Interaction Domain ◽  
Dihydroflavonol Reductase ◽  
Anthocyanin Biosynthetic Pathway ◽  
R2r3 Myb

The N-terminal of Myc-like basic helix-loop-helix transcription factors (bHLH TFs) contains an interaction domain, namely the MYB-interacting region (MIR), which interacts with the R2R3-MYB proteins to regulate genes involved in the anthocyanin biosynthetic pathway. However, the functions of MIR-domain bHLHs in this pathway are not fully understood. In this study, PbbHLH2 containing the MIR-domain was identified and its function investigated. The overexpression of PbbHLH2 in ”Zaosu” pear peel increased the anthocyanin content and the expression levels of late biosynthetic genes. Bimolecular fluorescence complementation showed that PbbHLH2 interacted with R2R3-MYB TFs PbMYB9, 10, and 10b in onion epidermal cells and confirmed that MIR-domain plays important roles in the interaction between the MIR-domain bHLH and R2R3-MYB TFs. Moreover, PbbHLH2 bound and activated the dihydroflavonol reductase promoter in yeast one-hybrid (Y1H) and dual-luciferase assays. Taken together these results suggested that the MIR domain of PbbHLH2 regulated anthocyanin biosynthesis in pear fruit peel.

An R2R3-MYB transcription factor CmMYB21 represses anthocyanin biosynthesis in color fading petals of chrysanthemum

2022 ◽  
Vol 293 ◽  
pp. 110674
Author(s):  
Yiguang Wang ◽  
Li-Jie Zhou ◽  
Yuxi Wang ◽  
Zhiqiang Geng ◽  
Baoqing Ding ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Anthocyanin Biosynthesis ◽  
Myb Transcription Factor ◽  
Color Fading ◽  
R2r3 Myb

scholarly journals A R2R3-MYB Transcription Factor, VvMYBC2L2, Functions as a Transcriptional Repressor of Anthocyanin Biosynthesis in Grapevine (Vitis vinifera L.)

Molecules ◽  
2018 ◽  
Vol 24 (1) ◽  
pp. 92 ◽  
Author(s):  
Ziguo Zhu ◽  
Guirong Li ◽  
Li Liu ◽  
Qingtian Zhang ◽  
Zhen Han ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Vitis Vinifera ◽  
Anthocyanin Biosynthesis ◽  
Transcriptional Repressor ◽  
Negative Regulator ◽  
Myb Transcription Factor ◽  
Vitis Vinifera L ◽  
Flavonoid Pathway ◽  
Leucoanthocyanidin Reductase ◽  
R2r3 Myb

In grapevine, the MYB transcription factors play an important role in the flavonoid pathway. Here, a R2R3-MYB transcription factor, VvMYBC2L2, isolated from Vitis vinifera cultivar Yatomi Rose, may be involved in anthocyanin biosynthesis as a transcriptional repressor. VvMYBC2L2 was shown to be a nuclear protein. The gene was shown to be strongly expressed in root, flower and seed tissue, but weakly expressed during the fruit development in grapevine. Overexpressing the VvMYBC2L2 gene in tobacco resulted in a very marked decrease in petal anthocyanin concentration. Expression analysis of flavonoid biosynthesis structural genes revealed that chalcone synthase (CHS), dihydroflavonol 4-reductase (DFR), leucoanthocyanidin reductase (LAR) and UDP glucose flavonoid 3-O-glucosyl transferase (UFGT) were strongly down-regulated in the VvMYBC2L2-overexpressed tobacco. In addition, transcription of the regulatory genes AN1a and AN1b was completely suppressed in transgenic plants. These results suggested that VvMYBC2L2 plays a role as a negative regulator of anthocyanin biosynthesis.

scholarly journals CsMYB3 and CsRuby1 form an ‘Activator-and-Repressor’ Loop for the Regulation of Anthocyanin Biosynthesis in Citrus

2019 ◽  
Vol 61 (2) ◽  
pp. 318-330 ◽  
Author(s):  
Ding Huang ◽  
Zhouzhou Tang ◽  
Jialing Fu ◽  
Yue Yuan ◽  
Xiuxin Deng ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Anthocyanin Biosynthesis ◽  
Myb Transcription Factor ◽  
Nitrogen Stress ◽  
Loop Model ◽  
Anthocyanin Accumulation ◽  
Promoter Activation ◽  
R2r3 Myb ◽  
Activation Capacity ◽  
Anthocyanin Biosynthetic Genes

Abstract Anthocyanins are preferentially accumulated in certain tissues of particular species of citrus. A R2R3-MYB transcription factor (named Ruby1) has been well documented as an activator of citrus anthocyanin biosynthesis. In this study, we characterized CsMYB3, a transcriptional repressor that regulates anthocyanin biosynthesis in citrus. CsMYB3 was expressed in anthocyanin-pigmented tissues, and the expression was closely associated with that of Ruby1, which is a key anthocyanin activator. Overexpression of CsMYB3 in Arabidopsis resulted in a decrease in anthocyanins under nitrogen stress. Overexpression of CsMYB3 in the background of CsRuby1-overexpressing strawberry and Arabidopsis reduced the anthocyanin accumulation level. Transient promoter activation assays revealed that CsMYB3 could repress the activation capacity of the complex formed by CsRuby1/CsbHLH1 for the anthocyanin biosynthetic genes. Moreover, CsMYB3 could be transcriptionally activated by CsRuby1 via promoter binding, thus forming an ‘activator-and-repressor’ loop to regulate anthocyanin biosynthesis in citrus. This study shows that CsMYB3 plays a repressor role in the regulation of anthocyanin biosynthesis and proposes an ‘activator-and-repressor’ loop model constituted by CsRuby1 and CsMYB3 in the regulation of anthocyanin biosynthesis in citrus.

scholarly journals The Paeonia qiui R2R3-MYB Transcription Factor PqMYB113 Positively Regulates Anthocyanin Accumulation in Arabidopsis thaliana and Tobacco

2022 ◽  
Vol 12 ◽  
Author(s):  
Xiaokun Liu ◽  
Jingjing Duan ◽  
Dan Huo ◽  
Qinqin Li ◽  
Qiaoyun Wang ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Bimolecular Fluorescence Complementation ◽  
Luciferase Reporter ◽  
Myb Transcription Factor ◽  
Anthocyanin Synthesis ◽  
Regulation Mechanism ◽  
Reporter Assay ◽  
Anthocyanin Accumulation ◽  
Tree Peony ◽  
R2r3 Myb

Paeonia qiui is a wild species of tree peony native to China. Its leaves are purplish red from the bud germination to the flowering stage, and anthocyanin is the main pigment in purplish red leaves. However, the anthocyanin synthesis regulation mechanism in tree peony leaves remains unclear. In this study, an R2R3-MYB, PqMYB113 was identified from the leaves of P. qiui. Phylogenetic analysis revealed that PqMYB113 clustered with Liquidambar LfMYB113 and grape VvMYBA6. Subcellular location analysis showed that PqMYB113 was located in the cell nucleus. The transient reporter assay suggested that PqMYB113 was a transcriptional activator. The overexpression of PqMYB113 in Arabidopsis thaliana and tobacco (Nicotiana tabacum) resulted in increased anthocyanin accumulation and the upregulation of CHS, F3H, F3’H, DFR, and ANS. The dual luciferase reporter assay showed that PqMYB113 could activate the promoters of PqDFR and PqANS. Bimolecular fluorescence complementation assays and yeast two-hybrid assays suggested that PqMYB113 could form a ternary MBW complex with PqbHLH1 and PqWD40 cofactors. These results provide insight into the regulation of anthocyanin biosynthesis in tree peony leaves.

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